US2024360403A1PendingUtilityA1

Transgene expression of ligninolytic enzymes in fungi

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Assignee: BATTELLE MEMORIAL INSTITUTEPriority: Apr 28, 2023Filed: Apr 29, 2024Published: Oct 31, 2024
Est. expiryApr 28, 2043(~16.8 yrs left)· nominal 20-yr term from priority
Inventors:Ziyu DaiAna L. RoblesJoonhoon KimScott E. BakerJon K. MagnusonBeth A. HofstadJohn M. Gladden
C12N 2501/998C08H 6/00C12Y 111/01016C12N 2500/20C12N 9/0061C12R 2001/645C12N 1/14C12N 9/0065C12Y 111/01014C12Y 111/01013C12N 15/80
67
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Claims

Abstract

Fungus with incorporated of heterologous ligninolytic enzyme to enhance effective conversion of lignocellulosic feedstocks to various chemical productions.

Claims

exact text as granted — not AI-modified
What is claimed: 
     
         1 . A modified filamentous fungus transformed with at least two polynucleotides coding for manganese peroxidase (mnP), versatile peroxidase, laccase or lignin peroxidase (lip). 
     
     
         2 . The fungus of  claim 1 , wherein the fungus is transformed with polynucleotides coding for manganese peroxidase (mnP) and laccase. 
     
     
         3 . The fungus of  claim 1 , wherein the fungus is transformed with polynucleotides coding for manganese peroxidase (mnP) and lip. 
     
     
         4 . The fungus of  claim 1 , wherein the fungus is transformed with polynucleotides coding for manganese peroxidase (mnP), laccase, versatile peroxidase and lip. 
     
     
         5 . The fungus of  claim 1 , wherein the polynucleotide comprises at least one intron. 
     
     
         6 . The fungus of  claim 1 , further comprising a disruption in expression of one or more of transcriptional factor of protease (prtT), subunit of Goli mannoyltransferase complex (mnn9), and V-SNARE binding protein (vsm1) genes. 
     
     
         7 . The fungus of  claim 6 , wherein expression of prtT gene is disrupted. 
     
     
         8 . The fungus of  claim 6 , wherein expression of prtT and vsm1 genes is disrupted. 
     
     
         9 . The fungus of  claim 6 , wherein the fungus is transformed with the manganese peroxidase polynucleotide and the prtT gene is disrupted. 
     
     
         10 . A method of culturing the modified fugus of  claim 1 , comprising culturing the fungus in culture medium comprising a protein selected from hemoglobin, soy protein, skim milk protein, serum albumin or combination thereof. 
     
     
         11 . The method of  claim 10 , further comprising culturing said fungus at a pH of about 4 to about 5. 
     
     
         12 . The method of  claim 10 , further comprising adding manganese to the culture medium. 
     
     
         13 . A method to increase expression of ligninolytic enzyme from a filamentous fungus comprising transforming said fungus with at least two polynucleotides encoding manganese peroxidase (mnP), versatile peroxidase, laccase or lignin peroxidase (lip) and culturing said transformed fungus so as to express at least two of manganese peroxidase (mnP), versatile peroxidase, laccase or lignin peroxidase (lip). 
     
     
         14 . The method of  claim 13 , wherein the fungus is transformed with manganese peroxidase (mnP) and laccase polynucleotides. 
     
     
         15 . The method of  claim 13 , wherein the fungus is transformed with manganese peroxidase (mnP) and lip polynucleotides. 
     
     
         16 . The method of  claim 14 , wherein the fungus is transformed with manganese peroxidase (mnP), laccase, versatile peroxidase and lip polynucleotides. 
     
     
         17 . The method of  claim 13 , wherein the polynucleotide comprises at least one intron. 
     
     
         18 . The method of  claim 13 , further comprising a disruption in expression of one or more of transcriptional factor of protease (prtT), subunit of Goli mannoyltransferase complex (mnn9), and V-SNARE binding protein (vsm1) genes. 
     
     
         19 . The method of  claim 18 , wherein expression of prtT gene is disrupted. 
     
     
         20 . The method of  claim 18 , wherein expression of prtT and vsm1 genes is disrupted. 
     
     
         21 . The method of  claim 18 , wherein the fungus is transformed with the manganese peroxidase polynucleotide and the prtT gene is disrupted. 
     
     
         22 . The method of  13  comprising, further culturing said transformed fungus in culture medium comprising a protein selected from hemoglobin, soy protein, skim milk protein, serum albumin or combination thereof. 
     
     
         23 . The method of  claim 13 , further comprising culturing said fungus at a pH of about 4 to about 5. 
     
     
         24 . The method of  claim 13 , further comprising adding manganese to the culture medium. 
     
     
         25 . The method of  claim 13 , further comprising harvesting one or more of manganese peroxidase (mnP), versatile peroxidase, laccase or lignin peroxidase (lip). 
     
     
         26 . A method to degrade lignin comprising contacting said lignin with the fungus of  claim 1  or one or more enzymes harvested therefrom.

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